"
User:DavidC/29 September 2009
From 2009.igem.org
(New page: {{Template:Supbiotechcss6.css}} {{Template:SupbiotechparisEn}} <center> <div style="color: black; margin-bottom: 30px; width: 500px;"> {|align="center" ||{{#calendar: title=User:DavidC |...) |
|||
| (2 intermediate revisions not shown) | |||
| Line 12: | Line 12: | ||
</div> | </div> | ||
</center> | </center> | ||
| + | |||
| + | |||
| + | === Tuesday the 29th === | ||
| + | |||
| + | ==== Ligation between D protein and adenovirus 5 penton base ==== | ||
| + | |||
| + | Restriction digest of D protein by Spe I (380bp): <br> | ||
| + | DNA (PCR with bio-brick primers) = 5µL <br> | ||
| + | Buffer M (TAKARA) =2µL <br> | ||
| + | BSA (TAKARA) = 2µL <br | ||
| + | H20 = 10µL <br> | ||
| + | Spe 1 (TAKARA) = 1µL <br> | ||
| + | 1 hour of incubation at 37°C. <br><br> | ||
| + | |||
| + | Restriction digest of adenovirus 5 penton base by XbaI (1715bp): <br> | ||
| + | |||
| + | DNA (PCR with bio-brick primers) = 5µL <br> | ||
| + | Buffer M (TAKARA) =2µL <br> | ||
| + | BSA (TAKARA) = 2µL <br | ||
| + | H20 = 10µL <br> | ||
| + | Xba 1 (TAKARA) = 1µL <br> | ||
| + | 1 hour of incubation at 37°C. <br> | ||
| + | |||
| + | ==== DNA electrophoresis ==== | ||
| + | |||
| + | 85 Volt, 15 minutes. <br> | ||
| + | 105 Volt, 40 minutes. <br> | ||
| + | Ladder fermentas 1 Kb. <br> | ||
| + | |||
| + | |||
| + | Samples: D protein cut by SpeI (D prt), ADV5pb protein cut by XbaI. | ||
| + | <br> | ||
| + | |||
| + | [[image:TU2909.png|center]] | ||
| + | <br> | ||
| + | |||
| + | ==== DNA purification ==== | ||
| + | |||
| + | Kit Qiagen “gel extraction kit”, final volume = 50µL. <br> | ||
Latest revision as of 15:15, 21 October 2009
Introduction Current therapies Gene therapies DVS Concept Tissue Vector Cell Vector Therapeutic plasmid Conclusion Implementation Tissue targeting Antitumor action Treatment modeling Conclusion
BioBricks Material & Method Bibliography
Contents |
Tuesday the 29th
Ligation between D protein and adenovirus 5 penton base
Restriction digest of D protein by Spe I (380bp):
DNA (PCR with bio-brick primers) = 5µL
Buffer M (TAKARA) =2µL
BSA (TAKARA) = 2µL <br
H20 = 10µL
Spe 1 (TAKARA) = 1µL
1 hour of incubation at 37°C.
Restriction digest of adenovirus 5 penton base by XbaI (1715bp):
DNA (PCR with bio-brick primers) = 5µL
Buffer M (TAKARA) =2µL
BSA (TAKARA) = 2µL <br
H20 = 10µL
Xba 1 (TAKARA) = 1µL
1 hour of incubation at 37°C.
DNA electrophoresis
85 Volt, 15 minutes.
105 Volt, 40 minutes.
Ladder fermentas 1 Kb.
Samples: D protein cut by SpeI (D prt), ADV5pb protein cut by XbaI.
DNA purification
Kit Qiagen “gel extraction kit”, final volume = 50µL.
